Detecting in-solution conformational changes in viral fusogens using tryptophan-induced fluorescence quenching

STAR Protoc. 2021 Dec 6;2(4):100994. doi: 10.1016/j.xpro.2021.100994. eCollection 2021 Dec 17.

Abstract

Dynamic monitoring of protein conformational changes is necessary to fully understand many biological processes. For example, viral entry and membrane fusion require rearrangement of its viral glycoprotein. We present a step-by-step protocol for site-specific bimane labeling of the influenza-C fusogen to map proximity and conformational movements using tryptophan-induced fluorescence quenching. This protocol is adaptable for other proteins and for protein-protein interaction detection. For complete details on the use and execution of this protocol, please refer to Serrão et al., 2021.

Keywords: Biophysics; Microbiology; Molecular/Chemical Probes; Protein Biochemistry; Protein expression and purification; Structural Biology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Gammainfluenzavirus / chemistry
  • Glycoproteins / analysis
  • Glycoproteins / chemistry
  • Glycoproteins / metabolism
  • Protein Conformation
  • Spectrometry, Fluorescence / methods*
  • Tryptophan / chemistry*
  • Tryptophan / metabolism
  • Viral Fusion Proteins* / analysis
  • Viral Fusion Proteins* / chemistry
  • Viral Fusion Proteins* / metabolism
  • Virus Internalization

Substances

  • Glycoproteins
  • Viral Fusion Proteins
  • Tryptophan